EP0141547A1
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Improved heterogeneous immunoassay method and assembly
Assignee: Baxter Healthcare Corp
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Filing: 1984-10-09
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Granted: —
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Expires: 2004-10-09
Abstract
Abstract A method and assembly (10) provided for the detection and determination of at least one ligand in a sample suspected of containing the ligand. The method includes providing in a hand-held assembly having a first chamber (14) and a second chamber (26) separated by a separation means (18) which permits the passage therethrough of materials of a relatively small size while preventing the passage therethrough of materials of a relatively larger size; a suspension (16) in the first chamber of an admixture of a binding partner for the ligand and a labeled component. The labeled component is selected from the group consisting of binding partner, compound which can bind to the binding partner, compound which can bind to the reaction product of the ligand and binding partner, and compound which can bind to the ligand. The sample suspected of containing the ligand is introduced to the first chamber. The mixture is incubated for a time sufficient to form a reaction mixture containing free labeled component and a complex of the labeled component, binding partner and ligand. The free labeled component is separated from the complex by creating pressure differential across the separation medium whereby the relatively larger size complex remains in the first chamber while the relatively smaller size free labeled component passes to the second chamber. The presence of either the labeled component present in the complex in the first chamber or the free labeled component in the second chamber is determined. The determination of such labeled component in either chamber provides a measure of the presence and quantity of the ligand in the sample.
Inventors
- 👤 Linda L. Jones
- 👤 Edward T. Maggio
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-04 18:09
CPC Classifications
B; B01; B01L; B01L3/00; B01L3/50; B01L3/502; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/536; G01N33/537
Claims Analysis
Total Claims: 51
Claim 1
Independent Claim
249 words
A method for the detection and determination of at least one ligand in a sample suspected of containing such Ugand, the method comprising the steps of: suspending in a hand-held assembly having at least a first chamber and a second chamber separated by a separation means for permitting the passage therethrough of materials of a relatively small size while preventing the passage therethrough of materials of a relatively larger size, an admixture in the first chamber comprising a binding partner for such ligand and a labeled component selected from the group consisting of binding partner, compound which can bind to the binding partner, compound which can bind to the reaction product of the ligand and binding partner, and compound which can bind to the ligand; introducing to such first chamber a sample suspected of containing such ligand; incubating the resultant mixture for a time sufficient to form a reaction mixture containing free labeled component and a complex of the labeled component, binding partner, and ligand; separating the complex from the free labeled component by creating a pressure differential across the separation means whereby the relative larger size complex remains in the first chamber while the relatively smaller size free labeled component passes to the second chamber; and determining the presence of either the labeled component present in the complex in the first chamber or the free labeled component in the second chamber, either of which comprises a measure of the presence and quantity of the ligand in the sample.
Claim 2
Independent Claim
249 words
A method for the detection and determination of at least one ligand in a sample suspected of containing such Ugand, the method comprising the steps of: suspending in a hand-held assembly having at least a first chamber and a second chamber separated by a separation means for permitting the passage therethrough of materials of a relatively small size while preventing the passage therethrough of materials of a relatively larger size, an admixture in the first chamber comprising a binding partner for such ligand and a labeled component selected from the group consisting of binding partner, compound which can bind to the binding partner, compound which can bind to the reaction product of the ligand and binding partner, and compound which can bind to the ligand; introducing to such first chamber a sample suspected of containing such ligand; incubating the resultant mixture for a time sufficient to form a reaction mixture containing free labeled component and a complex of the labeled component, binding partner, and ligand; separating the complex from the free labeled component by creating a pressure differential across the separation means whereby the relative larger size complex remains in the first chamber while the relatively smaller size free labeled component passes to the second chamber; and determining the presence of either the labeled component present in the complex in the first chamber or the free labeled component in the second chamber, either of which comprises a measure of the presence and quantity of the ligand in the sample.
Claim 3
Dependent Claim
30 words
A method as recited in claim 1 wherein the labeled component comprises a known concentration of a compound which competes with the ligand for binding sites on such binding partner.
Claim 4
Dependent Claim
18 words
A method as recited in claim 1 wherein the labeled component eomprises a known concentration of labeled ligand.
Claim 5
Dependent Claim
18 words
A method as recited in claim 1 wherein the labeled component comprises labeled binding partner for such ligand.
Claim 6
Dependent Claim
42 words
A method as recited in claim 1 further comprising the step of: binding a macromolecule to a compound selected from the group consisting of ligand, binding partner and complex, for enhancing the relative size differential between the complex and free labeled component.
Claim 7
Dependent Claim
74 words
A method as recited in claim 1 further comprising the steps of: providing in the first chamber a second labeled component, which second labeled component is nonreactive and labeled with the same label as the first labeled component; and determining the presence of any labeled component in the first chamber and in the second chamber which comprises a measure of the presence and quantity of the ligand in the sample and a reference control
Claim 8
Dependent Claim
101 words
A method as recited in claim 1 further comprising the steps of: providing at least one additional labeled component labeled with a different label than any other labeled component present in such first chamber and at least one additional binding partner in such first chamber, which such additional binding partner is capable of reacting with a different ligand which can be present in the sample; and deter mining-the presence of each of such separately labeled components in either the first chamber or the second chamber, which determinations comprise a measure of the presence and quantity of such ligands in the sample.
Claim 9
Dependent Claim
29 words
A method as recited in claim 1 wherein the reparation step comprises creating a pressure differential across the aeparation means by creating a reduced pressure in the second chamber.
Claim 10
Dependent Claim
55 words
A method as recited in claim 8 wherein the step of creating a reduced pressure comprises providing a collection vessel having an evacuated chamber as the second chamber and exposing the separation means to such evacuated chamber to cause the free labeled component to pass through the separation means and collect in the collection vesseL
Claim 11
Dependent Claim
29 words
A method as recited in claim 1 wherein the separation step comprises creating a pressure differential across the separation means by creating an increased pressure in the first chamber.
Claim 12
Dependent Claim
78 words
A method as recited in claim 10 wherein the step of creating an increased pressure comprises providing in such first chamber a collection vessel having a collection chamber as the second chamber and to which is attached the separation means, which collection vessel is movable with respect to the first chamber; in such first chamber to create such increased pressure and cause the free labeled component to pass through the separation means and collect in the collection vessel
Claim 13
Dependent Claim
20 words
A method as recited in claim 1 wherein the label determining step comprises providing label-determining reagent in the second chamber.
Claim 14
Dependent Claim
22 words
A method as recited in claim 12 wherein the label-determining reagent is provided in the second chamber prior to the separation step.
Claim 15
Dependent Claim
29 words
A method as recited in claim 12 wherein the labeled component is labeled with an enzyme and the label-determining reagent comprises a substrate and color agent for such enzyme.
Claim 16
Dependent Claim
26 words
A method as recited in claim 1 wherein the ligand to be determined comprises an antigen and the binding partner for such antigen comprises an antibody.
Claim 17
Dependent Claim
19 words
A method as recited in claim 15 wherein the labeled component comprises an enzyme labeled antibody for such antigen.
Claim 18
Dependent Claim
15 words
A method as recited in claim 15 wherein a macromolecule is bound to the antibody.
Claim 19
Dependent Claim
35 words
A method as recited in claim 1 wherein the suspending step comprises suspending an initially dry admixture of binding partner and labeled component by the step of introducing a suspending agent to the first chamber.
Claim 20
Dependent Claim
28 words
A method as recited in claim 18 wherein the suspending step comprises adding the sample to the first chamber as at least a portion of the suspending agent.
Claim 21
Dependent Claim
190 words
A hand-held assembly for the detection and determination of at least one ligand in a sample suspected of containing such ligand, the assembly comprising: a vessel having a first chamber for receiving a sample suspected of containing the ligand, which first chamber includes an admixture of a binding partner for such ligand and a labeled component selected from the group consisting of binding partner, compound which can bind to the binding partner, compound which can bind to the reaction product of the ligand and binding partner, and compound which can bind to the ligand; a second vessel having a second chamber separated from the first chamber by a separation means for permitting the passage therethrough of free labeled component of a relatively small size while preventing the passage therethrough of a complex formed from ligand, binding partner and labeled component of a relatively larger size than the free labeled component, when a pressure differential is created across such separation means; and means for creating a pressure differential across the separatior. means to separate the free labeled component from the complex and collect the free labeled component in the second chamber.
Claim 22
Dependent Claim
190 words
A hand-held assembly for the detection and determination of at least one ligand in a sample suspected of containing such ligand, the assembly comprising: a vessel having a first chamber for receiving a sample suspected of containing the ligand, which first chamber includes an admixture of a binding partner for such ligand and a labeled component selected from the group consisting of binding partner, compound which can bind to the binding partner, compound which can bind to the reaction product of the ligand and binding partner, and compound which can bind to the ligand; a second vessel having a second chamber separated from the first chamber by a separation means for permitting the passage therethrough of free labeled component of a relatively small size while preventing the passage therethrough of a complex formed from ligand, binding partner and labeled component of a relatively larger size than the free labeled component, when a pressure differential is created across such separation means; and means for creating a pressure differential across the separatior. means to separate the free labeled component from the complex and collect the free labeled component in the second chamber.
Claim 23
Dependent Claim
20 words
An assembly as recited in claim 20 wherein the admixture of binding partner and labeled component comprises a liquid suspension.
Claim 24
Dependent Claim
30 words
An assembly as recited in claim 20 wherein the labeled component comprises a known concentration of a compound which competes with the ligand for binding sites on such binding partner.
Claim 25
Dependent Claim
18 words
An assembly as recited in claim 20 wherein the labeled component comprises a known concentration of labeled ligand.
Claim 26
Dependent Claim
18 words
An assembly as recited in claim 20 wherein the labeled component comprises labeled binding partner for such ligand.
Claim 27
Dependent Claim
41 words
An assembly as recited in claim 20 further comprising a macromolecule in the admixture, which macromolecule comprises a compound capable of binding to a compound selected from the group consisting of ligand, binding partner and complex of ligand and binding partner.
Claim 28
Dependent Claim
33 words
An assembly as recited in claim 20 further comprising a second labeled component in the admixture, which second labeled component is non-reactive and labeled with the same label as the first labeled component.
Claim 29
Dependent Claim
69 words
An assembly as recited in claim 20 further comprising at least one additional labeled component in the admixture, which is labeled with a different label than any other labeled component present in the admixture in the first chamber; and at least one additional binding binding partner in such admixture, which such additional binding partner is capable of reacting with a different ligand which can be present in the sample.
Claim 30
Dependent Claim
16 words
An assembly as recited in claim 20 further comprising label detecting reagents in the second chamber.
Claim 31
Dependent Claim
102 words
An assembly as recited in claim 20 wherein the first vessel comprises: a generally cylindrical vessel defining the first chamber and. the separation means is positioned within the first chamber; an outlet port on the first vessel in fluid flow communication with the first chamber through such separation means; a needle hub means attached to the outlet port for retaining a needle cannula on the outlet port; a needle cannula attached to the needle hub means; and such second vessel comprises a generally cylindrically vessel defining a second chamber with a reduced pressure therein and a puneturable cover sealing the second chamber.
Claim 32
Dependent Claim
40 words
An assembly as recited in claim 29 further comprising a projecting wall portion on the first vessel, which projecting wall portion extends around the needle hub means and needle cannula and defines a cavity therewithin for receiving the second vessel
Claim 33
Dependent Claim
87 words
An assembly as recited in claim 20 wherein the first vessel comprises: a generally cylindrical vessel defining the first chamber therewithin and the second vessel comprises a generally cylindrical vessel removably positioned within the first chamber of the first vessel and wherein the separation means is positioned within the second vessel; and further comprising sealing means on the second vessel for providing a liquid seal between the second vessel and first vessel for preventing liquid from flowing into the annulus created between the first and second vessels.
Claim 34
Dependent Claim
19 words
An assembly as recited in claim 31 wherein the second chamber in the second vessel includes label detecting reagents.
Claim 35
Dependent Claim
71 words
An assembly as recited in claim 31 further comprising: a frangible bottom wall of the first chamber of the first veasel, which frangible bottom wall is selectively frangible upon a preselected pressure being exerted against such frangible bottom wall; and a third vessel attached to the first vessel, which third vessel defines a third chamber in communication with the first chamber upon fracturing the frangible bottom wall of the first vessel.
Claim 36
Dependent Claim
20 words
An assembly as recited in claim 33 further comprising label detecting reagents in the third chamber of the third vessel
Claim 37
Dependent Claim
19 words
An assembly as recited in claim 33 wherein the second chamber in the second vessel includes label detecting reagents.
Claim 38
Dependent Claim
267 words
A kit for the detection and determination of at least one ligand in a sample suspected of containing such ligand, the kit comprising: a first vessel having a first chamber for receiving a sample suspected of containing the ligand, which first chamber includes an admixture of a binding partner for such ligand and a labeled component selected from the group consisting of binding partner, compound which can bind to the binding partner, compound which can bind to the reaction product of the ligand and binding partner, and compound which can bind to the ligand; a separation means provided in the first vessel for permitting the passage therethrough of free labeled component of a relatively small size while preventing the passage therethrough of a complex formed from ligand, binding partner and labeled component of a relatively larger size than the free labeled component when a pressure differential is created across such separation means; a second vessel having a second chamber having a reduced pressure therein; a puneturable top enclosing the second chamber of the second vessel; an outlet port on the first vessel; a needle hub attached to the outlet port of the first vessel; and a needle cannula attached to the needle hub, which needle cannula is capable of piercing the top on the second vessel to expose the separation medium to the reduced pressure in the second vessel and create a pressure differential across the separation means for separating the free labeled component from the complex, which free labeled component flows through the needle eannula and is collected in the second chamber of the second vessel
Claim 39
Dependent Claim
267 words
A kit for the detection and determination of at least one ligand in a sample suspected of containing such ligand, the kit comprising: a first vessel having a first chamber for receiving a sample suspected of containing the ligand, which first chamber includes an admixture of a binding partner for such ligand and a labeled component selected from the group consisting of binding partner, compound which can bind to the binding partner, compound which can bind to the reaction product of the ligand and binding partner, and compound which can bind to the ligand; a separation means provided in the first vessel for permitting the passage therethrough of free labeled component of a relatively small size while preventing the passage therethrough of a complex formed from ligand, binding partner and labeled component of a relatively larger size than the free labeled component when a pressure differential is created across such separation means; a second vessel having a second chamber having a reduced pressure therein; a puneturable top enclosing the second chamber of the second vessel; an outlet port on the first vessel; a needle hub attached to the outlet port of the first vessel; and a needle cannula attached to the needle hub, which needle cannula is capable of piercing the top on the second vessel to expose the separation medium to the reduced pressure in the second vessel and create a pressure differential across the separation means for separating the free labeled component from the complex, which free labeled component flows through the needle eannula and is collected in the second chamber of the second vessel
Claim 40
Dependent Claim
49 words
A kit as recited in claim 36 further comprising a third vessel having a third chamber therein with a reduced pressure in the third chamber; a puncturable top on the third vessel sealing the third chamber, which puneturable top is puneturable by the needle cannula on the first vessel.
Claim 41
Dependent Claim
21 words
A kit as recited in claim 37 further comprising label detecting reagent present in the third chamber of the third vessel.
Claim 42
Dependent Claim
20 words
A kit as recited in claim 37 further comprising label detecting reagent in the second chamber of the second vesseL
Claim 43
Dependent Claim
35 words
A kit as recited in claim 36 further comprising a liquid suspending agent for introducing to the first chamber in the first vessel to provide a liquid suspension of the binding partner and labeled component.
Claim 44
Dependent Claim
28 words
A kit as recited in claim 36 wherein the admixture in the first chamber of the first vessel comprises a liquid suspension of binding partner and labeled component.
Claim 45
Dependent Claim
187 words
A kit for the detection and determination of at least one ligand in a sample suspected of containing such ligand, the kit comprising: a first vessel having a rust chamber for receiving a sample suspected of containing ligand, which first chamber includes an admixture of a binding partner for such ligand and a label component selected from the group consisting of binding partner, compound which can bind to the binding partner, compound which can bind to the reaction product of the ligand and binding partner, and compound which can bind to the ligand; a second vessel having a second chamber capable of being positioned within the first chamber of the first vessel, which second vessel includes sealing means for sealing the annulus between the second and first vessel to fluid flow; and separation means in the second chamber for permitting the passage therethrough of free labeled component of a relatively small size while preventing the passage therethrough of a complex formed from ligand, binding partner, and label component of a relatively larger than the free labeled component when a pressure differential is created across such separation means.
Claim 46
Dependent Claim
187 words
A kit for the detection and determination of at least one ligand in a sample suspected of containing such ligand, the kit comprising: a first vessel having a rust chamber for receiving a sample suspected of containing ligand, which first chamber includes an admixture of a binding partner for such ligand and a label component selected from the group consisting of binding partner, compound which can bind to the binding partner, compound which can bind to the reaction product of the ligand and binding partner, and compound which can bind to the ligand; a second vessel having a second chamber capable of being positioned within the first chamber of the first vessel, which second vessel includes sealing means for sealing the annulus between the second and first vessel to fluid flow; and separation means in the second chamber for permitting the passage therethrough of free labeled component of a relatively small size while preventing the passage therethrough of a complex formed from ligand, binding partner, and label component of a relatively larger than the free labeled component when a pressure differential is created across such separation means.
Claim 47
Dependent Claim
21 words
A kit as recited in claim 42 wherein the admixture comprises a liquid suspension of the binding partner and labeled component.
Claim 48
Dependent Claim
35 words
A kit as recited in claim 42 further comprising a liquid suspending agent for introducing to the first chamber of the first vessel to provide a liquid suspension of the binding partner and labeled component.
Claim 49
Dependent Claim
20 words
A kit as recited in claim 42 further comprising label detecting reagent in the second chamber of the second vessel.
Claim 50
Dependent Claim
32 words
A kit as recited in claim 42 further comprising a third vessel attachable to the first vesel and separated from the first vessel by a frangible bottom wall on the first vessel.
Claim 51
Dependent Claim
21 words
A kit aa recited in claim 46 further comprising label detecting reagent present in the third chamber of the third vessel.
References Cited (Backward Citations)
US4146365A
Cited
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